Application of conjoint liquid chromatography with monolithic disks for the simultaneous determination of immunoglobulin G and other proteins present in a cell culture medium.
The aim of this study was to develop a chromatographic method, as a substitute for enzyme-linked immunosorbent assays, for the rapid and simultaneous detection of IgG, insulin, and transferrin present in a cell culture medium. Conjoint liquid chromatography (conjoint LC) using monolithic disks was applied for this purpose. An anion-exchange disk was combined with a Protein G affinity disk in a preparative HPLC system. IgG bound to the Protein G disk, whereas transferrin and insulin were captured on the quaternary ammonium (QA) disk. Using this method, it was possible to simultaneously determine the concentrations of IgG, transferrin, and insulin in the cell culture medium. Thus, conjoint LC could be used for the rapid and simultaneous detection of different proteins present in a cell culture medium.
Ralla
Kathrin
K
Institute of Technical Chemistry, Gottfried Wilhelm Leibniz Universität Hannover, D-30167 Hanover, Germany.
Anton
Fabienne
F
Scheper
Thomas
T
Kasper
Cornelia
C
D000911:Antibodies, Monoclonal / Q000032:analysis*; D001426:Bacterial Proteins / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003470:Culture Media / Q000032:analysis; D007074:Immunoglobulin G / Q000032:analysis*; D007328:Insulin / Q000032:analysis*; D014168:Transferrin / Q000032:analysis*
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D003470:Culture Media; C034284:IgG Fc-binding protein, Streptococcus; D007074:Immunoglobulin G; D007328:Insulin; D014168:Transferrin